(from abstract). Recurrent herpes simplex virus type 1 (HSV-1) infection is a major cause of viral induced blindness. LAT, the only HSV-1 gene previously thought active during latency, is important for efficient reactivation of HSV-1. The PI has recently discovered a novel HSV-1 gene (AL) that, like LAT, is also transcriptionally active during latency, and is therefore a candidate for involvement in the latency-reactivation cycle. AL is antisense to LAT and overlaps the LAT promoter and the 5' end of LAT. The AL-RNA is polyadenylated and contains an ORF. Deletion of most of the AL-ORF and the corresponding region of LAT, produced a mutant with increased virulence and decreased spontaneous reactivation, suggesting that (1) AL and/or LAT are involved in spontaneous reactivation, and (2) AL-protein plays a role in virulence.
The specific aims i nclude: 1. Fine map and characterize the AL gene, RNA, and protein. Mapping will employ RT-PCR, RPA primer extension, RACE and cDNA cloning and sequencing, of HSV-1 RNA from tissue culture and rabbit trigeminal ganglia. Anti-AL protein antibody will be made and used to confirm the presence of an AL-protein. The time of AL expression will be determined. 2. Determine the function of AL. AL-/LAT+ mutants will be constructed and analyzed in rabbits for the effect on virulence and spontaneous reactivation. Since AL and LAT overlap, knocking out one gene without altering function of the other is complex. Approaches include: (a) single nucleotide changes in the AL-ORF ATG that knockout AL-protein without altering LAT RNA structure; (b) minimal LAT promoter alterations to block LAT transcription without altering the AL-protein amino acid sequence; and (c) expression of AL-protein from an alternative site in AL-/LAT+ and AL-/LAT- mutants. The construction and analysis of these complementary AL-/LAT+ and AL+/LAT- mutants should allow us to determine whether AL is involved in spontaneous reactivation and/or virulence.

National Institute of Health (NIH)
National Eye Institute (NEI)
Research Project (R01)
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Visual Sciences A Study Section (VISA)
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Shen, Grace L
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Cedars-Sinai Medical Center
Los Angeles
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Carpenter, Dale; Henderson, Gail; Hsiang, Chinhui et al. (2008) Introducing point mutations into the ATGs of the putative open reading frames of the HSV-1 gene encoding the latency associated transcript (LAT) reduces its anti-apoptosis activity. Microb Pathog 44:98-102
Carpenter, Dale; Hsiang, Chinhui; Brown, Donald J et al. (2007) Stable cell lines expressing high levels of the herpes simplex virus type 1 LAT are refractory to caspase 3 activation and DNA laddering following cold shock induced apoptosis. Virology 369:12-8
Jin, Ling; Perng, Guey-Chuen; Carpenter, Dale et al. (2007) Reactivation phenotype in rabbits of a herpes simplex virus type 1 mutant containing an unrelated antiapoptosis gene in place of latency-associated transcript. J Neurovirol 13:78-84
Nesburn, Anthony B; Bettahi, Ilham; Zhang, Xiuli et al. (2006) Topical/mucosal delivery of sub-unit vaccines that stimulate the ocular mucosal immune system. Ocul Surf 4:178-87
Jin, Ling; Perng, Guey-Chuen; Mott, Kevin R et al. (2005) A herpes simplex virus type 1 mutant expressing a baculovirus inhibitor of apoptosis gene in place of latency-associated transcript has a wild-type reactivation phenotype in the mouse. J Virol 79:12286-95
Barsam, Charles A; Brick, David J; Jones, Clinton et al. (2005) A viral model for corneal scarring and neovascularization following ocular infection of rabbits with a herpes simplex virus type 1 (HSV-1) mutant. Cornea 24:460-6
Peng, Weiping; Henderson, Gail; Inman, Melissa et al. (2005) The locus encompassing the latency-associated transcript of herpes simplex virus type 1 interferes with and delays interferon expression in productively infected neuroblastoma cells and trigeminal Ganglia of acutely infected mice. J Virol 79:6162-71
Jin, Ling; Perng, Guey-Chuen; Brick, David J et al. (2004) Methods for detecting the HSV-1 LAT anti-apoptosis activity in virus infected tissue culture cells. J Virol Methods 118:9-13
Henderson, Gail; Perng, Guey-Chuen; Nesburn, Anthony B et al. (2004) The latency-related gene encoded by bovine herpesvirus 1 can suppress caspase 3 and caspase 9 cleavage during productive infection. J Neurovirol 10:64-70
Peng, W; Jin, L; Henderson, G et al. (2004) Mapping herpes simplex virus type 1 latency-associated transcript sequences that protect from apoptosis mediated by a plasmid expressing caspase-8. J Neurovirol 10:260-5

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